Comparison of Hematopoietic Stem Cells Derived from Fresh and Cryopreserved Whole Cord Blood in the Generation of Humanized Mice

Scholbach, J.; Schulz, Anja; Westphal, Florian; Egger, Dietmar; Wege, Anja K.; Patties, Ina; Köberle, Margarethe; Sack, Ulrich; Lange, Franziska

doi:10.1371/journal.pone.0046772

Cited by 20 publications

(23 citation statements)

References 33 publications

(38 reference statements)

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“…Stable engraftment with human immune system consisting human lymphoid and myeloid cells was achieved in CD34-NOG-hIL-34 mice (Fig. 1e,f ), comparable to CD34-NSG (Additional file 1: Figure S3) [28][29][30][31]. Such human immune cell reconstitution levels are also similar with other existing humanized mouse models [32].…”

Section: Resultssupporting

confidence: 75%

Human Interleukin-34 facilitates microglia-like cell differentiation and persistent HIV-1 infection in humanized mice

Mathews

Woods

Katano

et al. 2019

Mol Neurodegeneration

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Background: Microglia are the principal innate immune defense cells of the centeral nervous system (CNS) and the target of the human immunodeficiency virus type one (HIV-1). A complete understanding of human microglial biology and function requires the cell's presence in a brain microenvironment. Lack of relevant animal models thus far has also precluded studies of HIV-1 infection. Productive viral infection in brain occurs only in human myeloid linage microglia and perivascular macrophages and requires cells present throughout the brain. Once infected, however, microglia become immune competent serving as sources of cellular neurotoxic factors leading to disrupted brain homeostasis and neurodegeneration.Methods: Herein, we created a humanized bone-marrow chimera producing human "microglia like" cells in NOD. Cg-Prkdc scid Il2rg tm1Sug Tg(CMV-IL34)1/Jic mice. Newborn mice were engrafted intrahepatically with umbilical cord blood derived CD34+ hematopoietic stem progenitor cells (HSPC). After 3 months of stable engraftment, animals were infected with HIV-1 ADA , a myeloid-specific tropic viral isolate. Virologic, immune and brain immunohistology were performed on blood, peripheral lymphoid tissues, and brain.Results: Human interleukin-34 under the control of the cytomegalovirus promoter inserted in NSG mouse strain drove brain reconstitution of HSPC derived peripheral macrophages into microglial-like cells. These human cells expressed canonical human microglial cell markers that included CD14, CD68, CD163, CD11b, ITGB2, CX3CR1, CSFR1, TREM2 and P2RY12. Prior restriction to HIV-1 infection in the rodent brain rested on an inability to reconstitute human microglia. Thus, the natural emergence of these cells from ingressed peripheral macrophages to the brain could allow, for the first time, the study of a CNS viral reservoir. To this end we monitored HIV-1 infection in a rodent brain. Viral RNA and HIV-1p24 antigens were readily observed in infected brain tissues. Deep RNA sequencing of these infected mice and differential expression analysis revealed human-specific molecular signatures representative of antiviral and neuroinflammatory responses.Conclusions: This humanized microglia mouse reflected human HIV-1 infection in its known principal reservoir and showed the development of disease-specific innate immune inflammatory and neurotoxic responses mirroring what can occur in an infected human brain.

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Section: Resultssupporting

confidence: 75%

Human Interleukin-34 facilitates microglia-like cell differentiation and persistent HIV-1 infection in humanized mice

Mathews

Woods

Katano

et al. 2019

Mol Neurodegeneration

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“…In contrast to previous reports in which cord blood progenitors were used to engraft neonatal NSG mice (Andre et al, 2010;McDermott et al, 2010;Choi et al, 2011b;Reimann et al, 2012;Scholbach et al, 2012;Singh et al, 2012), we detected very low levels (<0.5% CD45 + ) of human cells in the peripheral blood of most mice at 16 weeks after transplant (Fig. 1A).…”

Section: Durable Multilineage Engraftment In Adult Nsg Mice Transplancontrasting

confidence: 99%

Engraftment and Lineage Potential of Adult Hematopoietic Stem and Progenitor Cells Is Compromised Following Short-Term Culture in the Presence of an Aryl Hydrocarbon Receptor Antagonist

Torres-Coronado

Tran

et al. 2014

Human Gene Therapy Methods

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Hematopoietic stem cell gene therapy for HIV/AIDS is a promising alternative to lifelong antiretroviral therapy. One of the limitations of this approach is the number and quality of stem cells available for transplant following in vitro manipulations associated with stem cell isolation and genetic modification. The development of methods to increase the number of autologous, gene-modified stem cells available for transplantation would overcome this barrier. Hematopoietic stem and progenitor cells (HSPC) from adult growth factor-mobilized peripheral blood were cultured in the presence of an aryl hydrocarbon receptor antagonist (AhRA) previously shown to expand HSPC from umbilical cord blood. Qualitative and quantitative assessment of the hematopoietic potential of minimally cultured (MC-HSPC) or expanded HSPC (Exp-HSPC) was performed using an immunodeficient mouse model of transplantation. Our results demonstrate robust, multilineage engraftment of both MC-HSPC and Exp-HSPC although estimates of expansion based on stem cell phenotype were not supported by a corresponding increase in in vivo engrafting units. Bone marrow of animals transplanted with either MC-HSPC or Exp-HSPC contained secondary engrafting cells verifying the presence of primitive stem cells in both populations. However, the frequency of in vivo engrafting units among the more primitive CD34 + / CD90 + HSPC population was significantly lower in Exp-HSPC compared with MC-HSPC. Exp-HSPC also produced fewer lymphoid progeny and more myeloid progeny than MC-HSPC. These results reveal that in vitro culture of adult HSPC in AhRA maintains but does not increase the number of in vivo engrafting cells and that HSPC expanded in vitro contain defects in lymphopoiesis as assessed in this model system. Further investigation is required before implementation of this approach in the clinical setting.

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“…As noted above, some of the minor subsets of immune cells, such as Th17, may require that the recipient mouse expresses a human transgene. While BMC engraftments with cord blood are limited by the available abundance of CD34+ cells (10 4 to 10 5 per mouse), the greater abundance of fibular BMCs allows the transfer of 2 × 10 5 CD34+ cells (per mouse) to yield engraftment rates comparable to those with cord blood (Fu et al, 2017;Scholbach et al, 2012). Unlike the implantation of PDX into non-humanized mice, most if not all of the mice display human head and neck squamous cell carcinoma (HNSCC) tissue growing in subcutaneous tissue for the analysis.…”

Section: Understanding Resultsmentioning

confidence: 99%

Autologously Humanized Mice for Immune‐Oncologic Studies

Kim

2020

CP Pharmacology

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With the rapid approval of immune checkpoint inhibitors for lung, melanoma, breast, genitourinary, and hematological malignancies, the hematopoietic cells in the tumor microenvironment (TME) are now considered an important, if not essential, consideration for cancer scientists. In many instances, syngeneic murine models have not been highly predictive for responsiveness in clinical trials. Our limited understanding of the human TME have, therefore, precluded a rational translation of immunotherapeutic combinations. This has led to the adoption of hematopoietic humanized murine models for the study of human tumor immunology in vivo. However, concerns about chimerism rates, HLA mismatching, and incomplete reconstitution of the innate immune system have driven a quest for improvements in these allogeneic humanized murine systems. Presented in this article is a completely autologous xenotransplantation method for reconstituting the human tumor immune microenvironment in vivo without the use of a patient's peripheral blood which is known to be associated with low engraftment rates. With this new approach, the current limitations of allogeneic humanized models are avoided by using matched bone marrow cells (BMCs) and derived tumor xenoplants (PDXs) from solid tumors in cancer patients. This autologous system provides a platform for studying endogenous lymphocytic and myeloid cell infiltration into the human tumor in vivo.

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Comparison of Hematopoietic Stem Cells Derived from Fresh and Cryopreserved Whole Cord Blood in the Generation of Humanized Mice

Cited by 20 publications

References 33 publications

Human Interleukin-34 facilitates microglia-like cell differentiation and persistent HIV-1 infection in humanized mice

Human Interleukin-34 facilitates microglia-like cell differentiation and persistent HIV-1 infection in humanized mice

Engraftment and Lineage Potential of Adult Hematopoietic Stem and Progenitor Cells Is Compromised Following Short-Term Culture in the Presence of an Aryl Hydrocarbon Receptor Antagonist

Autologously Humanized Mice for Immune‐Oncologic Studies

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